Increased expression of CDK11p58 and cyclin D3 following spinal cord injury in rats
Increased expression of CDK11p58 and cyclin D3 following spinal cord injury in rats
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DOI：
10.1007/s11010-007-9642-z
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发表时间：
2008-02-01
影响因子：
4.3
通讯作者：
Shen, Aiguo
中科院分区：
文献类型：
--
作者：
Ji, Yuhong;Xiao, Feng;Shen, Aiguo
蛋白激酶是细胞正常生长发育的关键信号分子。CDK11(P58)是一种与p34(Cdc2)相关的蛋白激酶，在正常的细胞周期进程中起重要作用。然而，它在中枢神经系统(CNS)损伤中的分布和功能尚不清楚。本研究主要研究CDK11在脊髓损伤(SCI)过程中的蛋白表达和细胞定位。Western印迹分析显示CDK11(P58)在正常脊髓中未见表达。逐渐升高，在脊髓损伤后3d达高峰，然后下降。免疫组织化学方法进一步检测CDK11(P58)的蛋白表达。观察损伤后不同时间CDK11(P58)表达的变化。双重免疫荧光染色显示CDK11与NeuN、CNPase和GFAP共表达。CDK11/活性caspase-3和CDK11/增殖细胞核抗原在部分细胞中共定位。在正常和损伤脊髓中检测到与CDK11(P58)相关的细胞周期蛋白D3，并能增强其活性。在脊髓损伤过程中，细胞周期蛋白D3与CDK11(P58)的表达模式相似。免疫荧光双重染色显示CDK11与细胞周期蛋白D3在神经元和神经胶质细胞中共表达。免疫共沉淀进一步显示CDK11(P58)和细胞周期蛋白D3在损伤脊髓中相互作用。因此，CDK11(P58)和细胞周期蛋白D3在急性脊髓损伤后可能存在相互作用。CDK11(P58)的另一个伙伴是β-1，4-半乳糖基转移酶1(β-1，4-GT1)。免疫荧光分析显示CDK11/β-1，4-GT1在损伤脊髓中共定位。免疫共沉淀法检测到细胞周期蛋白D3-CDK4复合体的存在。综上所述，CDK11和细胞周期蛋白D3可能在脊髓病理生理过程中发挥重要作用。
Protein kinases are critical signalling molecules for normal cell growth and development. CDK11(p58) is a p34(cdc2)-related protein kinase, and plays an important role in normal cell cycle progression. However its distribution and function in the central nervous system (CNS) lesion remain unclear. In this study, we mainly investigated the protein expression and cellular localization of CDK11 during spinal cord injury (SCI). Western blot analysis revealed that CDK11(p58) was not detected in normal spinal cord. It gradually increased, reached a peak at 3 day after SCI, and then decreased. The protein expression of CDK11(p58) was further analyzed by immunohistochemistry. The variable immunostaining patterns of CDK11(p58) were visualized at different periods of injury. Double immunofluorescence staining showed that CDK11 was co-expressed with NeuN, CNPase and GFAP. Co-localization of CDK11/active caspase-3 and CDK11/proliferating cell nuclear antigen (PCNA) were detected in some cells. Cyclin D3, which was associated with CDK11(p58) and could enhance kinase activity, was detected in the normal and injured spinal cord. The cyclin D3 protein underwent a similar pattern with CDK11(p58) during SCI. Double immunofluorescence staining indicated that CDK11 co-expressed with cyclin D3 in neurons and glial cells. Coimmunoprecipitation further showed that CDK11(p58) and cyclin D3 interacted with each other in the damaged spinal cord. Thus, it is likely CDK11(p58) and cyclin D3 could interact with each other after acute SCI. Another partner of CDK11(p58) was beta-1,4-galactosyltransferase 1 (beta-1,4-GT 1). The co-localization of CDK11/beta-1,4-GT 1 in the damaged spinal cord was revealed by immunofluorescence analysis. The cyclin D3-CDK4 complexes were also present by coimmunoprecipitation analysis. Taken together, these data suggested that both CDK11 and cyclin D3 may play important roles in spinal cord pathophysiology.